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AB176648

NF kappaB p65 ELISA Kit

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(35 Publications)

NF kappaB p65 ELISA Kit is a single-wash 90-min Simplestep used to quantify NF kappaB p65 with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 30 citations

View Alternative Names

NFKB3, RELA, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3

5 Images
ELISA - NF kappaB p65 ELISA Kit (AB176648)
  • ELISA

Supplier Data

ELISA - NF kappaB p65 ELISA Kit (AB176648)
Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)

NFĸB p65 phosphorylation in response to TNFα treatment.

Induction of NFĸB p65 (pS536) phosphorylation in MCF-7 cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates, and treated (10 min) with a dose-range of TNFα before cell lysis. Data from quadruplicate measurements of NFĸB p65 (pS536) are plotted and compared against total NFĸB p65 protein levels. Comparative NFĸB p65 (pS536) and NFĸB p65 (Total) data also shown by Western Blot.

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)

Typical cell lysate dilution series.

Example of a typical NFĸB p65 cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)

Comparison of NFĸB p65 expression in different cell lines.

Cell line analysis for Total NFĸB p65 from 200 μg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 ELISA Kit (AB176648)

Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1 x Cell Extraction Buffer PTR. Data from duplicate measurements of NFĸB p65 (Total) are normalized and plotted.

Key facts

Detection method

Colorimetric

Sample types

Tissue Homogenate, Cell Lysate

Reacts with

Mouse, Human

Results type

Semi-Quantitative

Sensitivity

= 5 µg/mL

Range

6 - 600 µg/mL

Assay time

1h 30m

Assay Platform

Microplate

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

NF kappaB p65 ELISA Kit ab176648 is a rapid single-wash 90-min Other Immunoassay to measure NF kappaB p65 in cell lysate, tissue homogenate. This SimpleStep sensitivity is 5 µg/mL.

How the assay works

NF kappaB p65 SimpleStep ELISA®employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details.

Assay Specificity

Our SimpleStep ELISA® kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

NF kappaB p65 ELISA Kit ab176648 protocol summary

1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Precision

[ { "reproducibilityType": "Inter", "sample": "HeLa Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "8.7" }, { "reproducibilityType": "Intra", "sample": "HeLa Extract", "replicates": 6, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.8" } ]

What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

NF-kB p65 also known as RelA is a significant component of the NF-kB protein complex. This complex usually involves a molecular weight for p65 of approximately 65 kDa. NF-kB p65 is mechanistically a transcription factor that regulates genes involved in inflammation cell survival and immune response. Expression of p65 is widely seen in various cell types including immune cells and epithelial cells suggesting its role in numerous physiological processes.
Biological function summary

NF-kB p65 acts as part of the larger NF-kB complex usually forming a heterodimer with other family members like p50. This complex translocates to the nucleus upon activation where it binds specific DNA sequences to regulate gene expression. The p65 subunit is essential for transactivating target genes involved in immune and inflammatory responses. Its regulation is important for proper cellular functioning especially in the maintenance of immune homeostasis and inflammatory response.

Pathways

NF-kB p65 participates in critical pathways like the NF-kB signaling and Toll-like receptor pathways. These pathways are fundamental for initiating immune responses and contribute to the regulation of apoptosis and cellular stress responses. In the context of these pathways the IKK complex plays a pivotal role in NF-kB activation leading to the phosphorylation and subsequent degradation of inhibitors that retain NF-kB p65 in the cytoplasm therefore allowing its movement to the nucleus.

NF-kB p65 is implicated in conditions such as cancer and autoimmune diseases. Its dysregulation can lead to persistent activation of inflammatory pathways contributing to tumorigenesis and chronic inflammation. In the context of these diseases proteins such as the p50 subunit and the IKK complex are often involved reflecting the importance of tight regulation of NF-kB signaling in preventing pathological conditions.

Product protocols

Target data

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric RELA-NFKB1 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Besides its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (PubMed : 15790681). The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression. Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed : 33440148).
See full target information RELA

Publications (35)

Recent publications for all applications. Explore the full list and refine your search

eLife 13: PubMed39405093

2024

A new potential strategy for cutaneous squamous cell carcinoma treatment by generating serum-based antibodies from tumor-exposed mice.

Applications

Unspecified application

Species

Unspecified reactive species

Zheng Liu

Annals of medicine 55:2284366 PubMed37992411

2023

Elevated circulating inflammatory biomarker levels in the SIRT1-NF-κB-sCD40L pathway in patients with acute myocardial infarction: a case-control study.

Applications

Unspecified application

Species

Unspecified reactive species

Chunjuan Chen,Meiyi Zheng,Wei Wang,Wei Yu

Pathogens and global health 118:47-64 PubMed37978995

2023

Efficacy of artemether against toxocariasis in mice: parasitological and immunopathological changes in brain, liver, and lung.

Applications

Unspecified application

Species

Unspecified reactive species

Dina I Elgendy,Rasha A Elmahy,Alaa Ibrahim Mohamed Amer,Hoda A Ibrahim,Asmaa Fawzy Eltantawy,Fotouh Rashed Mansour,Amina M Salama

International journal of molecular sciences 24: PubMed38003411

2023

Bisphenol AF Induces Prostatic Dorsal Lobe Hyperplasia in Rats through Activation of the NF-κB Signaling Pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Sisi Huang,Kaiyue Wang,Dongyan Huang,Xin Su,Rongfu Yang,Congcong Shao,Juan Jiang,Jianhui Wu

Heliyon 9:e22055 PubMed38045213

2023

L. significantly alleviates cigarette smoke-induced acute lung injury by lowering NF-κB pathway activation and inflammatory mediators.

Applications

Unspecified application

Species

Unspecified reactive species

Nadia Hussain,Nadia Ikram,Kashif Ur Rehman Khan,Liaqat Hussain,Ali M Alqahtani,Taha Alqahtani,Musaddique Hussain,Muath Suliman,Mohammad Y Alshahrani,Basel Sitohy

Biomaterials research 27:65 PubMed37415210

2023

Oxygen-loaded microbubble-mediated sonoperfusion and oxygenation for neuroprotection after ischemic stroke reperfusion.

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Unspecified application

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Unspecified reactive species

Yi-Ju Ho,Hsiang-Lung Cheng,Lun-De Liao,Yu-Chun Lin,Hong-Chieh Tsai,Chih-Kuang Yeh

Redox biology 63:102714 PubMed37120929

2023

LPS-induced mitochondrial dysfunction regulates innate immunity activation and α-synuclein oligomerization in Parkinson's disease.

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Species

Unspecified reactive species

A Raquel Esteves,Diana F Silva,Diogo Banha,Emanuel Candeias,Beatriz Guedes,Sandra M Cardoso

Journal of physiology and pharmacology : an official journal of the Polish Physiological Society 73: PubMed37087565

2023

6-Shogaol protects against isoproterenol-induced cardiac injury in rats through attenutating oxidative stress, inflammation, apoptosis and activating nuclear respiratory factor-2/heme oxygenase-1 signaling pathway.

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Unspecified application

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Unspecified reactive species

H Li,J Shen,Y Zhang,L Hu,W Luo

BioFactors (Oxford, England) 49:699-711 PubMed36747328

2023

Crocetin regulates Th1/Th2 and Th17/Treg balances, nitric oxide production, and nuclear localization of NF-κB in Th2-provoked and normal situations in human-isolated lymphocytes.

Applications

Unspecified application

Species

Unspecified reactive species

Vafa Baradaran Rahimi,Saeideh Saadat,Pouria Rahmanian Devin,Ali Jebalbarezy,Maryam Moqaddam,Mohammad Hossein Boskabady,Vahid Reza Askari

Pharmaceuticals (Basel, Switzerland) 16: PubMed37259378

2023

Ganetespib with Methotrexate Acts Synergistically to Impede NF-κB/p65 Signaling in Human Lung Cancer A549 Cells.

Applications

Unspecified application

Species

Unspecified reactive species

Gehad Subaiea,Syed Mohd Danish Rizvi,Hemant Kumar Singh Yadav,Turki Al Hagbani,Marwa Helmy Abdallah,El-Sayed Khafagy,Hosahalli Veerabhadrappa Gangadharappa,Talib Hussain,Amr Selim Abu Lila
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